CORECEPTOR MODIFICATION OF TCR TYROSINE KINASE SIGNALS

TCR 酪氨酸激酶信号的辅助受体修饰

• 批准号: 2608130
• 负责人: M CARRIE MICELI
• 金额: $ 10.86万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-12-08 至 1999-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2608130
• 关键词: CD antigens; MHC class II antigen; T cell receptor; T lymphocyte; antigen presenting cell; apoptosis; biological signal transduction; cell growth regulation; gene mutation; hybridomas; interleukin 2; phosphorylation; protein tyrosine kinase; tissue /cell culture; transfection; western blottings

This proposal outlines studies on the functinal and enzymatic consequences of interplay between components of the T cell receptor (i.e., TCR alpha and beta chains CD3gamma, delta, epsilon and the zeta chain) and their associated protein tyrosine kinases (PTKs) and other T cell surface molecules and their associated PTK or phosphatase activities. Our working hypothesis is that the TCR can send a variety of enzymatically and functionally distinct signals by recruiting different cell surface "co-receptor" or "accesory" molecules, along with their associated PTK activities, into a TCR based signaling complex. The expression and arrangement of counter receptors on the antigen presenting cell (APC) would dictate which co-receptors and associated PTK actiities would be recruited for TCR-mediated signaling and where they would be positioned within the complex. In this manner, functionally distinct TCR-mediated signals dictated by the APC could be generate. The following specific aims are designed to test this hypothesis by expressing normal and altered forms of proteins involved in co- receptor/accessory molecule assisted TCR-mediated signal transduction in the class II-restricted antigen specific BI-141 T cell hybridoma and the class II expressing FT5.7 L cell APC and measuring the effects on TCR- mediated signal transduction. Specificaly, we propose to: 1) express mutant forms of p56lck alone or along with CD4 in the class II-restricted BI-141 T cell hybridoma and analyze TCR-mediated tyrosine phosphorylation and IL-2 producton, 2) analyze TCR-mediated cell suicide and inhibition of spontaneous gowth in these transfectants, 3) analyze the functional consequences of engaging different co-receptor/accessory molecules along with the TCR in BI-141 and transfectants expressing modified p56lck by stimulating them with APC transfected with individual counter-receptors, 4) analyze CD3-epsilon- and zeta chain- associated kinase activities and their relative contributions to specific signaling when different co- receptor or accessory molecules are included in the TCR signaling apparatus. These studie should contribute to our basic understanding of how TCR-mediated signals result in distinct functional consequences. Such an understanding may result in the development of interventive immune therapies designed to enhance the immune response to tumors, to efficiently vaccinate against tumors, or to stimulate T cell malignancies to undergo cell suicide. Furthermore, elucidation of TCR activation may help us understand how tumors escape immune destruction or what normal regulatory processes fail when a T cell is transformed into a leukemia or lymphoma.

该提案概述子功能和酶促研究 T细胞受体组分之间相互作用的结果 (i.e., TCR α和β链CD 3 γ、δ、β和ζ 链）及其相关蛋白酪氨酸激酶（PTK）和其他T 细胞表面分子及其相关的PTK或磷酸酶 活动 我们的工作假设是TCR可以发送各种各样的 在酶和功能上不同的信号， 不同的细胞表面“辅助受体”或“辅助”分子，沿着 其相关的PTK活性转化为基于TCR的信号传导复合物。 的 抗原递呈上反受体的表达和排列 细胞（APC）将决定哪些共受体和相关的PTK活性 将被招募用于TCR介导的信号传导， 位于综合体内。 以这种方式，功能上不同 可以产生由APC支配的TCR介导的信号。 的 下面的具体目标是为了测试这一假设， 表达正常和改变形式的蛋白质， 受体/辅助分子辅助的TCR介导的信号转导 II类限制性抗原特异性BI-141 T细胞杂交瘤和 II类表达FT 5.7 L细胞APC并测量对TCR-1的影响。 介导的信号转导。 具体而言，我们建议：1）表达 突变型p56 lck II类限制性BI-141 T细胞中单独或沿着CD 4 杂交瘤和分析TCR介导酪氨酸磷酸化和IL-2 2）分析TCR介导的细胞自杀和抑制 在这些转染子中的自发生长，3）分析功能性的 将不同的共受体/辅助分子沿着 用BI-141中的TCR和表达修饰的p56 lck的转染子， 用转染有单个反受体的APC刺激它们， 4)分析CD 3-β和ζ链相关激酶活性， 它们对特定信号传导的相对贡献， 受体或辅助分子包括在TCR信号传导中 设备. 这些研究应该有助于我们对 TCR介导的信号如何导致不同的功能后果。 这样的理解可能会导致干预性的发展。 免疫疗法旨在增强对肿瘤的免疫反应， 有效地接种抗肿瘤疫苗，或刺激T细胞恶性肿瘤 进行细胞自杀 此外，阐明TCR活化可 帮助我们了解肿瘤是如何逃脱免疫破坏的， 当T细胞转化为白血病时， 或者淋巴瘤